Polyprenyl phosphates have been well established as carriers in sugar transfer reactions involved in cell envelope polysaccharide biosynthesis in procaryotes and glycoprotein biosynthesis in eucaryotes. It is the objective of this research to study the mechanism and regulation of the biosynthesis of these polyprenyl phosphates in bacteria. Undecaprenyl pyrophosphate synthetase from Lactobacillus plantarum will be purified to homogeneity using in part hydrophobic affinity chromatography. The synthesis of several types of polyisoprenylated-Agarose will be carried out to test the application of hydrophobic column chromatography to the separation and analysis of several prenyl transferases. Undecaprenyl pyrophosphate synthesis will be characterized with regard to the number of polypeptide components, amino acid composition and peptide maps. A variety of allylic pyrophosphate substrate analogues will be tested with the synthetase to examine the properties of the enzyme's catalytic site and a proposed scheme for termination of product elongation. A number of site-specific radiolabeled compounds, including photoaffinity reagents and arginine specific reagents will be synthesized and reacted with the synthetase. Peptide maps of these modified enzymes will be made and interpreted to gain information about the substrate and product binding domains. Analogues of the pyrophosphoryl group of delta 3-isopentenyl pyrophosphate will be synthetized and tested as inhibitors of undecaprenyl pyrophosphate phosphatase. Two aspects of control will be studied. Studies of substrate binding constants using a variety of allylic substrates will be made using geranylgeranyl-, solanesyl-, and undecaprenyl pyrophosphate synthetases from Micrococcus luteus to analyze possible mechanisms of control of the utilization of common polyprenol precursors. A variety of nucleotide-sugars will be tested as activators of undecaprenyl pyrophosphate synthetase, undecaprenyl pyrophosphate phosphatase and undecaprenol kinase in L. plantarum and Staphylococcus aureus. These studies are important in understanding possible mechanisms of regulation of the concentration of undecaprenyl monophosphate in bacterial cells.